M.V. Moreno-Arribas

Capillary isoelectric focusing of erythropoietin glycoforms and its comparison with flat-bed isoelectric focusing and capillary zone electrophoresis

The influence of several operation conditions on separation of recombinant human erythropoietin glycoforms by capillary isoelectric focusing (cIEF) is explored. From this study it is deduced that in order to separate several glycoforms of erythropoietin, urea has to be added to sample, which should not be completely depleted of the excipients used in its formulation. On-line desalting does not provide separation enhancement for samples with high content of salt. Better resolution is obtained using a mixture of a broad and a narrow pH-range carrier ampholytes than with either one used separately. Under the experimental conditions, focusing voltages of 25 kV improve separation compared to lower and higher electric fields. Focusing times shorter than the time necessary for electric current to reach a minimum provide similar separations than longer focusing times at which a minimum value of the current has already been achieved. The optimized method allows the separation and quantitation in 12 min of at least seven bands containing glycoforms of recombinant erythropoietin with apparent isoelectric points in the range 3.78-4.69. Compared to flat-bed isoelectric focusing, cIEF provides better separation of bands of glycoforms in a shorter time, and allows quantitative determination. Capillary zone electrophoresis (CZE) gives rise to resolution of erythropoietin glycoforms similar to that obtained by cIEF. Although CZE requires a longer analysis time, its reproducibility in terms of peak area of glycoforms is better than in cIEF.

Antibacterial activity of wine phenolic compounds and oenological extracts against potential respiratory pathogens

Aims:  To investigate the effect of seven wine phenolic compounds and six oenological phenolic extracts on the growth of pathogenic bacteria associated with respiratory diseases (Pseudomonas aeruginosa, Staphylococcus aureus, Moraxella catarrhalis, Enterococcus faecalis, Streptococcus sp Group F, Streptococcus agalactiae and Streptococcus pneumoniae).

Analytical performance of three commonly used extraction methods for the gas chromatography-mass spectrometry analysis of wine volatile compounds.

The analytical performance of three extraction procedures based on cold liquid-liquid extraction using dicloromethane (LLE), solid phase extraction (SPE) using a styrene-divinylbenzene copolymer and headspace solid phase microextraction (SPME) using a carboxen-polydimethylsiloxane coated fibre has been evaluated based on the analysis of 30 representative wine volatile compounds. From the comparison of the three procedures, LLE and SPE showed very good linearity covering a wide range of concentrations of wine volatile compounds, low detection limits, high recovery for most of the volatile compounds under study and higher sensitivity compared to the headspace-SPME procedure. The latter showed in general, poor recovery for polar volatile compounds. Despite some drawbacks associated with the LLE and SPE procedures such as the more tedious sampling treatment and the use of organic solvents, the analytical performance of both procedures showed that they are more adequate for the analysis of wine volatiles.

Inactivation of oenological lactic acid bacteria (Lactobacillus hilgardii and Pediococcus pentosaceus) by wine phenolic compounds

Aims:  To investigate the inactivation properties of different classes of phenolic compounds present in wine against two wine isolates of Lactobacillus hilgardii and Pediococcus pentosaceus, and to explore their inactivation mechanism.

Impact of the nonvolatile wine matrix composition on the in vivo aroma release from wines.

The impact of the nonvolatile wine matrix composition on the retronasal aroma release of four volatile compounds added to different types of wines has been evaluated. For this purpose, a tailor-made retronasal aroma trapping device (RATD) was used to entrap the exhaled breath of six panelists previously trained in a specific consumption procedure. Five wines of different composition (white wine, sparkling white wine, young red wine, aged red wine, and a sweet wine) were evaluated. Prior to the evaluation, with the exception of the sweet wine, the wines were adjusted to the same ethanol content and aromatized with a mixture of four target volatile compounds. Aroma release data were submitted to multivariate statistical analysis in order to relate wine chemical composition and aroma release during wine drinking. Results showed interindividual differences and a clustering of panelists among lower and higher aroma releasers, which was in agreement to the differences in their breathing capacity. A significant influence of the matrix composition in the low aroma releasers group during wine consumption was observed. The consumption of red wines provoked a significantly higher aroma release than the consumption of white and sweet wines. From the chemical composition determined in the wine samples (pH, total acidity, total polyphenols, neutral polysaccharides, residual sugar, and nitrogenous compounds), the amount of total polyphenols was better correlated with the observed effect.

An Integrated View of the Effects of Wine Polyphenols and Their Relevant Metabolites on Gut and Host Health

Over the last few decades, polyphenols, and flavonoids in particular, have attracted the interest of researchers, as they have been associated with the health-promoting effects derived from diets rich in vegetables and fruits, including moderate wine consumption. Recent scientific evidence suggests that wine polyphenols exert their effects through interactions with the gut microbiota, as they seem to modulate microbiota and, at the same time, are metabolized by intestinal bacteria into specific bioavailable metabolites. Microbial metabolites are better absorbed than their precursors and may be responsible for positive health activities in the digestive system (local effects) and, after being absorbed, in tissues and organs (systemic effects). Differences in gut microbiota composition and functionality among individuals can affect polyphenol activity and, therefore, their health effects. The aim of this review is to integrate the understanding of the metabolism and mechanisms of action of wine polyphenols at both local and systemic levels, underlining their impact on the gut microbiome and the inter-individual variability associated with polyphenols’ metabolism and further physiological effects. The advent of promising dietary approaches linked to wine polyphenols beyond the gut microbiota community and metabolism are also discussed.

Microbial Contribution to Wine Aroma and Its Intended Use for Wine Quality Improvement

Wine is a complex matrix that includes components with different chemical natures, the volatile compounds being responsible for wine aroma quality. The microbial ecosystem of grapes and wine, including Saccharomyces and non-Saccharomyces yeasts, as well as lactic acid bacteria, is considered by winemakers and oenologists as a decisive factor influencing wine aroma and consumer’s preferences. The challenges and opportunities emanating from the contribution of wine microbiome to the production of high quality wines are astounding. This review focuses on the current knowledge about the impact of microorganisms in wine aroma and flavour, and the biochemical reactions and pathways in which they participate, therefore contributing to both the quality and acceptability of wine. In this context, an overview of genetic and transcriptional studies to explain and interpret these effects is included, and new directions are proposed. It also considers the contribution of human oral microbiota to wine aroma conversion and perception during wine consumption. The potential use of wine yeasts and lactic acid bacteria as biological tools to enhance wine quality and the advent of promising advice allowed by pioneering -omics technologies on wine research are also discussed.

Evolution of red wine anthocyanins during malolactic fermentation, postfermentative treatments and ageing with lees.

A comparative study was conducted on nine batches of wine, from the same initial wine, subjected to malolactic fermentation and ageing in barrels, under different technological conditions: Malolactic fermentation in barrel or in tank, with or without wine clarification, ageing with or without lees and stirring or no stirring of the lees. Samples were taken of the initial wine, of the wine at the end of malolactic fermentation, of the wines after clarifying treatments, and after 3, 6, 9, 12 and 14 months of ageing in the barrel, making a total of 48 wines. As a result of the anthocyanin analysis of all the wines studied, a total of 21 different anthocyanin compounds were detected, which can be classified into four groups: simple glucosides, acetyl glucosides, cinnamoyl glucosides and pyroanthocyanins. During MLF, it was shown that the effect of the container used seems to be more important than the metabolic activity of the bacteria responsible for the process. From application of the LSD test, significant differences were found in the concentrations of all the anthocyanin compounds identified due to ageing time and significant differences were also revealed for most anthocyanin compounds in relation to the manufacturing method, especially the presence or absence of lees.

Feasibility and application of liquid-liquid extraction combined with gas chromatography-mass spectrometry for the analysis of phenolic acids from grape polyphenols degraded by human faecal microbiota.

In this study the feasibility of a LLE-GC-EI-MS method for the analysis of 43 phenolic acids belonging to different chemical structure families which have been described in the literature as microbial-derived metabolites after consumption of dietary polyphenols was proved. In addition, the method was applied for the characterisation of phenolic metabolites resulting from the incubation, in anaerobic conditions, of a commercial grape seed extract (GSE) and their corresponding flavan-3-ol monomeric (GSE-M) and oligomeric (GSE-O) fractions with human faeces from healthy volunteers (n=3). The method showed average values of repeatability and reproducibility of 5.0% and 6.3%, respectively, adequate and low detection (1.8-30.8 μg L(-1)) and quantification limits (6.0-102.8 μg L(-1)) and good recovery values (95%, as average value). A total of 27 phenolic acids were identified in the faecal solutions after incubation with the grape seed extracts. In general, faecal samples incubated with GSE and GSE-M (monomeric fraction) yield a higher formation of phenolic acids compared to the samples incubated with the oligomer fraction (GSE-O).

Towards the Fecal Metabolome Derived from Moderate Red Wine Intake

Dietary polyphenols, including red wine phenolic compounds, are extensively metabolized during their passage through the gastrointestinal tract; and their biological effects at the gut level (i.e., anti-inflammatory activity, microbiota modulation, interaction with cells, among others) seem to be due more to their microbial-derived metabolites rather than to the original forms found in food. In an effort to improve our understanding of the biological effects that phenolic compounds exert at the gut level, this paper summarizes the changes observed in the human fecal metabolome after an intervention study consisting of a daily consumption of 250 mL of wine during four weeks by healthy volunteers (n = 33). It assembles data from two analytical approaches: (1) UPLC-ESI-MS/MS analysis of phenolic metabolites in fecal solutions (targeted analysis); and (2) UHPLC-TOF MS analysis of the fecal solutions (non-targeted analysis). Both approaches revealed statistically-significant changes in the concentration of several metabolites as a consequence of the wine intake. Similarity and complementarity between targeted and non-targeted approaches in the analysis of the fecal metabolome are discussed. Both strategies allowed the definition of a complex metabolic profile derived from wine intake. Likewise, the identification of endogenous markers could lead to new hypotheses to unravel the relationship between moderate wine consumption and the metabolic functionality of gut microbiota.